Anti-tumor n-nitrosomorpholine



United States Patent 3,275,517 ANTI-TUMOR N-NITROSOMORPHOLINE JeanBradley Harrison, Niagara Falls, N.Y., and Howard E. Hotfman,Wilmington, Del., assignors to E. I. du Pont de Nemours and Company,Wilmington, Del., a corporation of Delaware No Drawing. Filed Oct. 7,1964, Ser. No. 402,320 1 Claim. (Cl. 167-78) This application is acontinuation-in-part of our application Serial No. 265,373 filed March15, 1963, now abandoned, which is a continuation-in-part of our nowabandoned application Serial No. 156,157 filed November 30, 1961. p

This invention relates to inhibition of tumor growth. More particularlyit relates to inhibition of tumor growth using N-nitrosomorpholine.

According to this invention, N-nitrosomorpholine is an eflectivechemotherapeutic agent for the treatment of tumor growths. In laboratorytests on rats, intraperitoneal injections have been shown to delay orinhibit the regeneration of liver tissue surviving after a two-thirdspartial hepatectomy. This activity is extremely rare in chemicalcompounds and is characteristic, for example, ofN,N',N"-triethy1enephosphoramide, an anti-tumor agent of merit in somecancer treatments.

The N-nitrosomorpholine will be administered in a tumorgrowth-inhibiting amount. Doses of from 0.1 to about 75 milligrams perkilogram of body weight of the recipient can be used as needed.Administration can be orally, intraveneously, intraarterially,intraperitoneally, intracutaneously, subcutaneously, intramuscularly, byregional intraarterial infusion, percutaneously or interaiticularly.

Our discovery of the anti-tumor activity of N-nitrosomorpholine is ahighly useful and exciting research development. The compound has shownactivity in mice against a variety of tumors. It has shown particularlyhigh activity against the tumor Mecca lymphosarcoma. Other tumors thatcould be afiected include malignant adenoma, hemangioendothelioma,adenocarcinoma, myeloma, epidermoid carcinoma, leukemia, bronchiogeniccarcinoma, lymphosarcoma, malignant melanoma, reticulum cell sarcoma,retinoblastoma, leiomyosarcoma, osteogenic sarcoma, rhabdomyosarcoma,fibrosarcoma, teratoma, chondrosarcoma, endothelial sarcoma,lymphangioendot-helioma, and liposarcoma.

The following examples are provided to more clearly define ourinvention.

Example 1 A daily dose of 30 milligrams per kilogram of 'body weighteffects significant growth inhibition of the tumor Mecca lymphosarcom-ain a host mouse. The dosage, tumor and host can be varied, as will bereadily understood, without departing from the concept of the presentinvention.

Example 2 Sarcoma 180, ascitic form, was injected intraperitoneally intoten mice. The amount of tumor was approximately one million cells permouse. Starting 24 hours after administration of the tumor cells,injection of N-nitrosomorpholine was made intraperitoneally once eachday for seven days. At the end of ten days, the intraperitoneal fluidwas removed and compared to the weight of the fluid in the control mice.The resultant data obtained from tests at two dose levels ofN-nitrosomorpholine are given below.

3,275,517 Patented Sept. 27, 1966 "ice Tumor T/C Body Weight Deaths Dosegm. Change T/C T/C 5 1 mgJkg 4. 0/7. 9 +2. 0/7. 0 0/5 30 mg./kg e 6/9.3 1. 5/+8 0 l Aseites fluid: weight of fluid from treated mice/weight oifluid from control mice.

2 Body weight change oi treated mice/body weight change of controls. aNumber of deaths 0! treated mice/number of deaths of controls.

This data indicates the compound N-nitrosomorpholine significantlyinhibited the growth of the ascites tumor at 15 and 30 mg./kg.

Example 3 tumor, the animals are individually weighed to detect.

possible weight loss which might suggest intoxication.

After two weeks each tumor in an experimental group was measured in situby means of calipers through the largest apparent diameter and through asecond diameter perpendicular to it. The average tumor diameter of theexperimental group and of the control group were compared and thegrading, weight change and deaths noted. The data obtained are asfollows:

Tumor Size Body Weight Deaths Dose /C, cm. Change T/C, TIC

30 mg./kg 1. /2. 60 3. 5/+3. 0 0/5 A significant inhibition of the tumorgrowth was observed.

Example 4 Using the Ehrlich carcinoma, an experiment as described inExample 3 was completed in a similar manner. The data obtained using theEhrlich carcinoma are as follows:

Tumor Size Body Weight Deaths Dose T/C, cm. Change TIC, 0

30 mgJkg 1. 85/2. 60 I -3.5/ +3.0 I 015 A significant inhibition of thetumor growth was seen.

Example 5 Using the Lewis lung carcinoma, an experiment as described inExample 3 was completed in a similar manner. The data obtained using theLewis lung carcinoma are as follows:

Tumor Size Body Weight Deaths Dose T/C, cm. Change T/C, T/C

30 mgJkg 1. 13/1. 85 I 3. 0/+5. 0

These data indicate that a significant inhibition in tumor growth wasobserved.

the Mecca lymphosarcoma.

a rm t Using the Mecca lymphosarcoma, an expe 1 en as de The mventlonclalmed 1S:

scribed in Example 3 was completed in a similar manner with oneexception. The growth of the tumor was measured after three weeks. Thedata obtained using the Mecca lymphosarcoma are as follows:

5 comprising administering to said animal from about 0.1

to about 75 milligrams of N-nitrosomoljpholine per kilogram of bodyweight of the recipient.

References Cited by the Examiner Tumor Size Body weight Deaths 10Sugiura, Cancer Research, vol. 25, No. 3, part 2,Apri1 Dose T/C, cm.Change T/C, /O 1965, pp. 494 to 521.

i p JULIAN s. LEVITT, Primary Examiner. mmg'lkg H's/+12 JEROME D.GOLDBERG, Assistant Examiner.

Example 6 This data represents a highly significant inhibition of Amethod of inhibiting tumor growth in an animal l

